Feline Ubiquitin Fusion Protein Genes

Using cDNA from a CRFK cell line as a template, PCR amplification was performed with the Ub1S and poly(dT) primers to isolate feline ubiquitin genes. Sequencing of the 495 bp PCR fragment revealed that the putative amino acids induced by this fragment gave a fusion protein consisting of a ubiquitin polypeptide (76 amino acids) and an extension protein of ribosomal proteins L40 (52 amino acids). The putative amino acid sequence of ubiquitin was identical to those of humans, rats and pigs.The recombinant glutathione S-transferase (GST)–feline ubiquitin fusion proteins were produced in Escherichia coli and purified. The fusion proteins had a molecular weight of about 42 kDa and were detected by immunoblot assay with rabbit anti-ubiquitin antiserum.The mRNAs from heat-shocked and non-heat-shocked cells were subjected to RT-PCR (Ub1S and poly(dT) primers) analysis. The molecular weights of the ubiquitinated proteins in heat-shocked CFRK cells were between 18 kDa and 24 kDa by immunoblot assay.These results suggested that there were more ubiquinated proteins in the heat-shocked CRFK cells than in the pre-heat-shocked cells.     

The significance of the structural characteristics of the agent for the problems of immunization against blue tongue]

Bluetongue virus (BTV), an arthropod-borne virus, is transmitted primarily by biting midges of the genus Culicoides. Some insect species, which might serve as a potential vector, are prevalent in Central Europe. In sheep, bluetongue is acute and mortality is high, whereas in cattle, goats and most wild ruminants the infection is usually clinically inapparent. Viremia is of short duration in sheep, but cattle experience a prolonged viremia and provide a reservoir for the dissemination of BTV. At least 24 different BTV serotypes have been identified. Antigenic variations occur in the polypeptides of the outer viral capsid and the segmented nature of the viral genome provides the potential for evolution of the virus by a mechanism of reassortment. This renders the use of polyvalent vaccines inefficient and emphasizes the significance of import/export restrictions on ruminants from BTV endemic areas.     

Teratogenicity testing of BI 58 EC (38% dimethoate) in chicken embryos with special respect to degradation of the active ingredient.

The insecticide formulation BI 58 EC was tested for teratogenicity in chicken embryos, with particular reference to degradation of the active ingredient (dimethoate) after the treatment of embryonated eggs. The pesticide was diluted in water to a concentration level of 0.8%, and the emulsion was injected into the air space in a volume of 0.1 ml/egg, or hen's eggs were treated by the immersion technique. Residues of dimethoate were measured in the samples on days, 13, 15 and 19 of the incubation of chicken embryos, and morphological examinations were performed simultaneously. Analytical chemistry data indicated a slower degradation of dimethoate in embryos after the immersion of eggs, and cyllosis was remarkable in this group among the sporadic developmental anomalies. The liver tissues of both treated groups exhibited severe fatty infiltration.     

Characterisation of cross-reactivity of virus neutralising antibodies induced by feline panleukopenia virus and canine parvoviruses

It was recently reported that canine parvoviruses (CPV) had entered cat populations and induced disease in infected cats, while they had affected only dogs in the past. It is important to determine whether conventional feline panleukopenia virus (FPLV) vaccines protect against recent CPV infections. In this study, the cross-reactivity of virus-neutralising (VN) and haemagglutinin-inhibition (HI) antibodies in cats induced by FPLV and CPV s were examined. Lower cross-reactivities of VN and HI antibodies against each CPV strain were observed in cats experimentally inoculated with FPLV or vaccinated with an inactivated FPLV vaccine. In addition, we revealed the existence of a novel type of FPLV, which reacted weakly with antibodies induced by the conventional FPLV vaccine.     

Milk fever and subclinical hypocalcaemia--an evaluation of parameters on incidence risk, diagnosis, risk factors and biological effects as input for a decision support system for disease control.

The present review analyses the documentation on incidence, diagnosis, risk factors and effects of milk fever and subclinical hypocalcaemia. It is hereby evaluated whether the existing documentation seems sufficient for further modelling in a decision support system for selection of a control strategy. Several studies have been carried out revealing an incidence of milk fever most often in the level of 5-10%. Few studies indicate that the incidence of subclinical hypocalcaemia is several times higher than milk fever. The diagnosis based on clinical or laboratory methods or based on presence of risk factors is outlined. The clinical symptoms of milk fever are highly specific and the disease level may thus be determined from recording of treatments. Diagnosis of subclinical hypocalcaemia needs to include laboratory examinations or it may be determined by multiplying the incidence of milk fever by a certain factor. From the documentation on risk factors, it is very complex to predict the incidence from the exposure level of the risk factors. Due to uncertainty, sensitivity analyses over a wide range of values for each parameter are needed. The documentation of cow characteristics, nutrition, environment and management as risk factors are described. Among cow characteristics, parity or age, body condition and production level were found to be important. Risk factors associated with nutrition included most importantly dietary cation-anion difference and calcium level whereas the importance of general feeding related factors like type of feed stuff and feeding level were less clear. Environment and management included season, climate, housing, pasturing, exercise, length of dry period and prepartum milking. Several of the parameters on environment and management were confounded among each other and therefore firm conclusions on the importance were difficult. The documentation of the effect of milk fever includes the downer cows, reproductive disorders, occurrence of other diseases and the effect on milk production, body weight and culling. The reproductive disorders included most importantly dystocia, uterine prolapse, retained placenta, metritis and repeat breeding, and occurrence of other diseases included ketosis, displaced abomasum and mastitis. The documentation was substantial and often quantifiable within certain limits. Overall it is concluded that the present documentation on milk fever concerning incidence, diagnosis, risk factors and effects seems sufficient for a systematic inclusion in a decision support system. A model on milk fever should take into consideration the variation in biological data and individual herd characteristics. The inclusion of subclinical hypocalcaemia would be more uncertain and probably should await further documentation on possibilities of determining the herd level incidence and also the effect of this condition on production.     

A review of diet formulation strategies and feeding systems to reduce excretory and feed wastes in aquaculture

Significant efforts should be devoted to reducing waste outputs from aquaculture operations in order to lower the environmental impacts of aquaculture in many parts of the world. Since most aquaculture wastes are ultimately from dietary origin, reduction of waste outputs should first be through improvements of diet formulation and feeding strategies. The first step in the production of feeds producing less solid waste is to eliminate poorly digestible ingredients (such as whole grain or grain by-products used as binders and fillers in the feed formulae) and to use highly digestible ingredients with good binding properties. Further reduction of solid waste can then be achieved through careful selection of the ingredients to improve apparent digestibility and the nutrient balance of the feed. Nitrogen waste outputs can be reduced through the reduction of the digestible protein to digestible energy (DP/DE) ratio of the diet. Phosphorus waste outputs can be reduced through careful selection of the ingredients and optimization of the digestible phosphorus content of the diet to meet the requirement of the fish but avoid greatly exceeding this required level. Finally, feeding practices that minimize feed wastage should be adopted since feed wastage can have a very significant impact on waste outputs from fish culture operations.     

Monthly model for genetic evaluation of laying hens 1. Fixed regression.

1. This paper addresses the possibility of using a monthly model for the genetic evaluation of laying hens, based on the definition of a test day model with fixed regression as used in dairy cattle, in which monthly records were treated as repeated measurements of the same trait. 2. Production records of 6450 hens, daughters of 180 sires and 1335 dams were analysed using an animal model with restricted maximum likelihood (REML). The traits considered were individual monthly egg production and cumulative egg production in 11 months. Four different models were fitted to various combinations of monthly and cumulative records. The covariates were derived from the regression of Ali and Schaeffer (1987). 3. Spearman rank correlations were computed to compare breeding values from different models. Two types of correlations were computed: between individual breeding values and between sire breeding values based on subsets of full-sib records. 4. The results indicated that a monthly model with nested covariates produced higher heritability and permanent environmental variance than the models with non-nested or without covariates. The estimates of heritability obtained from monthly model were lower than the estimates from the cumulative model. The monthly model resulted in higher correlations of sire breeding values between two subsets of full-sib records than those from cumulative models. 5. In conclusion, the monthly model with nested covariates appears to be better than the model with non-nested covariates or without covariate. Although the heritability estimates obtained from the monthly model were lower, the monthly model with nested covariates could be better than the cumulative model for genetic evaluation of laying hens in the 1st cycle of laying period when using either full or part records. The use of information from odd months of production could be of interest for the evaluation of full records.     

Localization of extracellular matrix receptors in ICGN mice, a strain of mice with hereditary nephrotic syndrome.

Fibrotic degeneration was examined in the kidneys of ICR-derived glomerulonephritis (ICGN) mice, a novel inbred mouse line with a hereditary nephrotic syndrome of unknown etiology considered to be a good model of human idiopathic nephrotic syndrome. In the present study, we histochemically revealed changes in accumulation of extracellular matrix (ECM) components and in localization of integrins, cellular receptors for ECM, in the kidneys of ICGN mice with the progression of renal failure. Excessive accumulation of basement membrane (laminin and collagen IV) and interstitial (type III collagen) ECM components were demonstrated in the glomeruli and tubulointerstitum of ICGN mice. Marked deposition of type I collagen and tenascin was seen only in the glomeruli of ICGN mice but not in those of ICR mice as normal controls. Increased expression of integrin alpha1-, alpha2-, alpha5- and beta1-subunits in glomeruli with fibrotic degeneration and abnormal distribution of alpha6-subunit were noted in the kidneys of ICGN mice. Excessive laminin, a ligand of alpha6beta1-integrin, was demonstrated on the tubular basement membrane, but alpha6-subunit diffusely disappeared on the basal side of the tubular epithelial cells. We presumed that abnormal integrin expression in renal tubules causes epithelial cell detachment, and consequently tubular nephropathy, and results in disorder of ECM metabolism causing excessive accumulation of ECM components in the kidneys of ICGN mice.     

Protection of turbot, Scophthalmus maximus (L.), and rainbow trout, Oncorhynchus mykiss (Richardson), against vibriosis using two different vaccines

Abstact. The potency of a whole-cell bacterin (WCB) and a toxoid enriched whole-cell vaccine (WCEB) administered intraperitoncally into rainbow trout, Oncorhynchus mykiss (Richardson), were compared. The most effective vaccine was further evaluated by bathing turbot, Scophthalmus maximus (L.). These vaccines were composed of three strains of V. anguillarun , of the serotypes 01 and 02. Both vaccines conferred the highest protection against strains of serotype 01 within 4 weeks. With the toxoid enriched vaccine giving the best results (77 RPS). When trout were revaccinated after 7 weeks with this vaccine, good protection was achieved against strains of serotypes 01 and 02. Interestingly, when the WCEB was administered by bath to turbot, acceptable levels of protection against strains of both serotypes were obtained after 4 weeks of immunization.